LOCUS       sequence-428684-        2789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synthetic terminator for gene expression in yeasts and fungi. Part 
            type 4 following the YeastToolkit MoClo grammar..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2789)
  AUTHORS   Hebra T, Smrckova H, Elkatmis B, Prevorovsky M, Pluskal T
  TITLE     POMBOX: A Fission Yeast Cloning Toolkit for Molecular and Synthetic 
            Biology.
  JOURNAL   ACS Synth Biol. 2023 Nov 22. doi: 10.1021/acssynbio.3c00529.
  PUBMED    37991801
REFERENCE   2  (bases 1 to 2789)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2023 Nov 22. doi: 10.1021/acssynbio.3c00529."
COMMENT     Sequence Label: pPOM029_P4_tSynth25
FEATURES             Location/Qualifiers
     source          1..2789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..119
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     313..335
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     327..344
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     328..344
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(568..584)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(568..584)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(581..603)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    592..608
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(616..622,623..640,641..646))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    661..682
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(799..816)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(970..1558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1050..1069)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1729..2520,2521..2589))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2352..2371
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2590..2694)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2762..2780
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
       61 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      121 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      181 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      241 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      301 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg gagatcggta
      361 cttcgcgaat gcgtcgagat ctagacatcc gtctcatcgg tctcaatcct aactcgagtt
      421 tttttttata tatatatata tatatatata actgtctaga aataaagagt atcatctttc
      481 aaagctgtga gacctgagac gcatatgcac atcggatgcc gggaccgacg agtgcagagg
      541 cgtgcaagcg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      601 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      661 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      721 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      781 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      841 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      901 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      961 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1021 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1081 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1141 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1201 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1261 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1321 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1381 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1441 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1501 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1561 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1621 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1681 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1741 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1801 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1861 gataccgcga gatccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1921 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1981 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2041 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2101 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2161 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2221 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2281 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2341 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2401 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2461 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2521 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2581 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2641 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2701 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     2761 aaataggcgt atcacgaggc cctttcgtc
//
